Vitamin B-related Gene Polymorphisms and Cardiovascular Disease.

Hyperhomocysteinemia is an independent risk factor for atherosclerosis, even in early childhood. A mutation in genes that code homocysteine metabolism enzymes or deficiency of specific vitamin cofactors may cause hyperhomocysteinemia. Vitamin B complex has been correlated with serum homocysteine levels. Any abnormality in its metabolism or nutritional deficiency may lead to hyperhomocysteinemia. Both vitamin B complex and homocysteine levels are partly genetically determined. Specifically, the most studied polymorphism is 677T-C in exon 5 of the 5,10- methylenetetrahydrofolate reductase (MTHFR) gene, which plays an important role in folate's metabolism. This polymorphism has been shown to be correlated with hypertension and cardiovascular disease. Polymorphisms in methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 1-like (MTHFD1L) gene have also been correlated with increased risk for coronary artery disease. Other common serious polymorphisms regard the area with high linkage disequilibrium, including the neuroblastoma breakpoint family, NBPF3 gene, and ~ 12-50 kb upstream of the tissue nonspecific alkaline phosphatase gene. Finally, the polymorphisms which have been mostly associated with vitamin B12 concentration are the rs11254363 polymorphism at intron 52 of the intrinsic factor vitamin B12 receptor of the CUBN and the rs526934 polymorphism at intron 8 of transcobalamin I. To sum up, several polymorphisms have already been associated with vitamin B complexes and therefore homocysteine level, highlighting the complex nature of vitamin B genetics.

Nursing staff fatigue and burnout during the COVID-19 pandemic in Greece.

INTRODUCTION: The coronavirus pandemic (COVID-19) is an unprecedented global health crisis with emotional and physical impact on health care workers. OBJECTIVE: The purpose of this study was to investigate the levels of fatigue and burnout in nursing staff during the pandemic. METHODS: The present study involved nursing staff from hospitals in Greece in February 2021, who completed the Fatigue (FAS) and Burnout (CBI) questionnaires. Gender, age, years of work experience, workplace (COVID-19 or non-COVID-19 wards) and SARS-CoV-2 infection status were recorded. RESULTS: The sample included 593 women and 108 men, with a mean age ± SD: 42.9 ± 9.9 years and 18.14 ± 10.8 years work experience. Slightly more than half, (367, 52.4%) worked in COVID-19 departments. Fifty-six (8%) tested positive for SARS-CoV-2 and 14 of them needed to be treated. The mean ± SD FAS and CBI scores were 25.6 ± 7.4 and 46.9 ± 18.8, respectively (67.9% and 42.9% had scores suggestive of fatigue and burnout, respectively). Women showed higher values in both scales (p < 0.01). Subjects working in COVID-19 wards scored significantly higher on both the FAS and CBI scales; they were also younger and with less work experience (p < 0.01). Staff treated for COVID-19 scored higher on the burnout scale (p < 0.01) than the uninfected staff. Fatigue showed a strong positive correlation with burnout (p < 0.01, r = 0.70). Stepwise multiple regression showed that the variation of fatigue was explained by 47.0% and 6.1% by the scores on the subscales of personal and work-related burnout, respectively. CONCLUSION: In conclusion, high rates of fatigue and burnout were found in the studied population. Nurses working with COVID-19 patients had higher rates of fatigue and burnout compared to those working elsewhere. There was a strong positive correlation (r = 0.70) between burnout and fatigue. Particular attention should be paid to staff who became ill and need to be treated.

Characterization of the emerging B.1.621 variant of interest of SARS-CoV-2

The genetic diversity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has the potential to impact the virus transmissibility and the escape from natural infection- or vaccine-elicited neutralizing antibodies. Here, representative samples from circulating SARS-CoV-2 in Colombia between January and April 2021, were processed for genome sequencing and lineage determination following the nanopore amplicon ARTIC network protocol and PANGOLIN pipeline. This strategy allowed us to identify the emergence of the B.1.621 lineage, considered a variant of interest (VOI) with the accumulation of several substitutions affecting the Spike protein, including the amino acid changes T95I, Y144T, Y145S and the insertion 146N in the N-terminal domain, R346K, E484K and N501Y in the Receptor-binding Domain (RBD) and P681H1 in the S1/S2 cleavage site of the Spike protein. The rapid increase in frequency and fixation in a relatively short time in Magdalena, Atlantico, Bolivar, Bogota D.C, and Santander that were near the theoretical herd immunity suggests an epidemiologic impact. Further studies will be required to assess the biological and epidemiologic roles of the substitution pattern found in the B.1.621 lineage. HighlightsO_LIMonitoring the emergence of new variants of SARS-CoV-2 in real time is a worldwide priority. C_LIO_LIEmerging variants of SARS-CoV-2 may have high impact biological implications for public health C_LIO_LIThe SARS-CoV-2 B.1.621 variant of interest was characterized by several substitutions: T95I, Y144T, Y145S, ins146N, R346K, E484K, N501Y and P681H in spike protein. C_LI

The roles of triglyceride/high-density lipoprotein cholesterol ratio and uric acid as predisposing factors for metabolic syndrome in healthy children

PURPOSE: To estimate the roles of triglyceride/high-density lipoprotein cholesterol (TG/HDL) ratio and uric acid in predisposition for metabolic syndrome (MetS) and its components in healthy children. METHODS: Anthropometric and biochemical analyses were performed on 110 children, aged 5 to 12 years, from the Greek county of Laconia. The children were studied as a whole population and in separate groups according to age and predisposition to MetS after taking into consideration International Diabetes Federation criteria, body mass index, and lipid profile. RESULTS: Seventeen percent of children exhibited predisposition to MetS, while 39.1% had TG/HDL ratio >1, and 3.64% had high level of uric acid. According to a receiver operating characteristic curve analysis, the relative probability for MetS predisposition sextupled when TG/HDL ratio was ≥1 (odds ratio [OR], 5.986; 95% confidence interval [CI], 1.968–18.205). Children in the total population and those aged < 9 years had a greater probability for increased low-density lipoprotein (LDL) cholesterol (OR, 3.614; 95% CI, 1.561–8.365) when TG/HDL ratio was ≥ 1. The TG/HDL ratio was positively correlated with body mass index (BMI) (P=0.035) in children without MetS, cholesterol in the total population (P=0.06) and children ≥9 years old (P=0.026), and with LDL in the total population and both age groups (P=0.001). The TG/HDL ratio was also positively correlated with alanine aminotransferase in the total population (P=0.033) and gamma-glutamyl transferase in most studied groups (P<0.001). Uric acid was positively correlated with waist circumference in the total population (P=0.043) and in those without MetS (P=0.027). It was also positively correlated with BMI, TG, cholesterol, and TG/HDL ratio and negatively correlated with HDL in most studied groups (P<0.005). CONCLUSION: The studied parameters correlated with MetS components and could be characterized as effective indexes for childhood MetS, regardless of age and predisposition to MetS.

The role of p21/CIP1/WAF1 (p21) in the negative regulation of the growth hormone/growth hormone receptor and epidermal growth factor/epidermal growth factor receptor pathways, in growth hormone transduction defect

PURPOSE: Growth hormone transduction defect (GHTD) is characterized by severe short stature, impaired STAT3 (signal transducer and activator of transcription-3) phosphorylation and overexpression of the cytokine inducible SH2 containing protein (CIS) and p21/CIP1/WAF1. To investigate the role of p21/CIP1/WAF1 in the negative regulation of the growth hormone (GH)/GH receptor and Epidermal Growth Factor (EGF)/EGF Receptor pathways in GHTD. METHODS: Fibroblast cultures were developed from gingival biopsies of 1 GHTD patient and 1 control. The protein expression and the cellular localization of p21/CIP1/WAF1 was studied by Western immunoblotting and immunofluorescence, respectively: at the basal state and after induction with 200-μg/L human GH (hGH) (GH200), either with or without siRNA CIS (siCIS); at the basal state and after inductions with 200-μg/L hGH (GH200), 1,000-μg/L hGH (GH1000) or 50-ng/mL EGF. RESULTS: After GH200/siCIS, the protein expression and nuclear localization of p21 were reduced in the patient. After successful induction of GH signaling (control, GH200; patient, GH1000), the protein expression and nuclear localization of p21 were reduced. After induction with EGF, p21 translocated to the cytoplasm in the control, whereas in the GHTD patient it remained located in the nucleus. CONCLUSIONS: In the GHTD fibroblasts, when CIS is reduced, either after siCIS or after a higher dose of hGH (GH1000), p21’s antiproliferative effect (nuclear localization) is also reduced and GH signaling is activated. There also appears to be a positive relationship between the 2 inhibitors of GH signaling, CIS and p21. Finally, in GHTD, p21 seems to participate in the regulation of both the GH and EGF/EGFR pathways, depending upon its cellular location.

Prevalence of Brucella antibodies on a previously acute brucellosis infected population: sensitivity, specificity and predictive values of Rose Bengal and Wright standard tube agglutination tests.

PURPOSE: Brucellosis is a zoonosis with worldwide distribution. The presence of antibodies after acute infection and the prevalence of positive serology in endemic area are not well documented. METHODS: Patients hospitalized with acute brucellosis were relocated 3-13 years after the initial infection. Hospital records of the initial infection were retrieved, and examination of Brucella antibodies using Rose Bengal test (RBT) and Wright standard tube agglutination (STA) test was performed. RESULTS: Eighty-three patients were hospitalized from 2000 to 2010; 50.6 % were farmers and 37.4 % livestock farmers. All had febrile illness and various focal complications. All had positive serology, and 82.2 % had positive blood cultures; 91.5 % were treated with streptomycin plus doxycycline. Seventy-two (86.7 %) were relocated on follow-up. Nine (12.5 %) had positive RBT and STA up to 1/320. Occupational history was associated with positive serology (p = 0.0172), and 8/9 of the positive individuals were livestock farmers (38.0 % of the livestock farmers checked). Residence, years after the infection, clinical presentation of brucellosis and treatment were not associated with serology results. Both tests had excellent sensitivity (nearly 100 %), specificity 87.5 % and excellent negative predictive value (nearly 100 %); however, positive predictive value was only 11.4 %. CONCLUSIONS: Rapid and low-cost tests as RBT and STA are still very useful in diagnosing acute brucellosis; however, every positive test must be examined together with clinical symptoms and occupational history. The tests can be used as screening tests in endemic populations to rule out acute brucellosis.

Protective mechanisms against oxidative stress and angiopathy in young patients with diabetes type 1 (DM1).

OBJECTIVE: Advanced glycation end-products (AGEs) via their receptor, RAGE, are involved in diabetic angiopathy. Soluble RAGE, an inhibitor of this axis, is formed by enzymatic catalysis (sRAGE) or alternative splicing (esRAGE). Malondialdehyde (MDA) is an oxidative stress marker, and ferric reducing ability of plasma (FRAP) is an anti-oxidant capacity marker. METHODS: In isolated mononuclear blood cells from 110 DM1-patients (P) and 124 controls (C) (4-29 years) RAGE mRNA (g) and protein expression (pe) were measured by RT-PCR and Western immunoblotting, respectively. Plasma levels of CML (AGEs) and sRAGE were measured by ELISA, MDA by flurometry and FRAP according to 'Benzie and Strain'. RESULTS: P showed: (i) higher g of RAGE, especially in p>13 years of age and >5 years DM1, (ii) increased pe of esRAGE in DM1>5 years and (iii) increased FRAP and MDA. CONCLUSIONS: The increased esRAGE and FRAP with increased levels of CML and MDA possibly reflects a protective response against the formation of diabetic complications in these young diabetic patients.

Increased growth hormone receptor (GHR) degradation due to over-expression of cytokine inducible SH2 domain-containing protein (CIS) as a cause of GH transduction defect (GHTD).

OBJECTIVE: Children with growth hormone transduction defect (GHTD) have impaired growth and signal transducer and activator of transcription 3 (STAT3) activation. Here, we examine the etiology of GHTD. METHODS: Control (Cf) and GHTD (Pf) children's fibroblasts were induced with hGH, MG132, lactacystin or silence RNA/CIS (siCIS). Western immunoblotting (WI) examined protein expression. Immunofluorescent microscopy (IF) examined cellular localization. RESULTS: (i) Pf showed retarded activation of pJAK2 and pSTAT-5 and increased ubiquitinated CIS (UbCIS) by WI. (ii) After MG132, Pf showed normal activation of pJAK2, pSTAT5 and pSTAT3. (iii) IF showed membrane (ML) and cytoplasmic localization (CL) of the GHR in Cf while the Pf showed only CL. In Pf, induction with lactacystin or siCIS changed the localization of the GHR to ML. CONCLUSIONS: In GHTD, abnormal GH signalling may be caused by over-expression of CIS, which may increase degradation of GHR, thus reducing membrane GHR availability, delaying activation of pJAK2 and pSTAT5 and reducing activation of pSTAT3.

Association between serum levels of the soluble receptor (sRAGE) for advanced glycation endproducts (AGEs) and their receptor (RAGE) in peripheral blood mononuclear cells of children with type 1 diabetes mellitus.

AIM: The binding of AGEs to RAGE is involved in diabetic vascular complications. We studied sRAGE levels and RAGE protein expression (P) together with N-carboxymethyl lysine (CML), a major AGE, in 74 patients with type 1 diabetes mellitus (DM1) and 43 healthy (C) children. METHODS: sRAGE and CML levels were determined by ELISA and RAGE P was evaluated in mononuclear cells by Western immunoblotting. RESULTS: Serum sRAGE was higher in DM1 than in C (1430 +/- 759 vs 1158 +/- 595 pg/ml, p = 0.047), inversely correlated to diabetes duration (r = -0.265, p = 0.037) and directly correlated to LDL-cholesterol levels (r = 0.224, p = 0.039). Diabetes duration correlated independently with sRAGE (p = 0.034). Circulating CML levels were not significantly different between DM1 and C groups (3.51 +/- 1.49 vs 3.59 +/- 1.83 ng/ml, p > 0.05) and RAGE P was lower in DM1 than in C (61 +/- 46 vs 102 +/- 63%, p = 0.0001). CONCLUSIONS: Increased serum sRAGE in children with DM1 may provide temporary protection against cell damage and may be sufficient to eliminate excessive circulating CML.

The bone-specific transcriptional regulator Cbfa1 is a target of mechanical signals in osteoblastic cells.

A primary goal of bone research is to understand the mechanism(s) by which mechanical forces dictate the cellular and metabolic activities of osteoblasts, the bone-forming cells. Several studies indicate that osteblastic cells respond to physical loading by transducing signals that alter gene expression patterns. Accumulated data have documented the fundamental role of the osteoblast-specific transcription factor Cbfa1 (core-binding factor) in osteoblast differentiation and function. Here, we demonstrate that low level mechanical deformation (stretching) of human osteoblastic cells directly up-regulates the expression and DNA binding activity of Cbfa1. This effect seems to be fine tuned by stretch-triggered induction of distinct mitogen-activated protein kinase cascades. Our novel finding that activated extracellular signal-regulated kinase mitogen-activated protein kinase physically interacts and phosphorylates endogenous Cbfa1 in vivo (ultimately potentiating this transcription factor) provides a molecular link between mechanostressing and stimulation of osteoblast differentiation. Elucidation of the specific modifiers and cofactors that operate in this mechanotranscription circuitry will contribute to a better understanding of mechanical load-induced bone formation which may set the basis for nonpharmacological intervention in bone loss pathologies.